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Quantification and Whole Genome Characterization of SARS-CoV-2 RNA in Wastewater and Air Samples
Published on: June 30, 2023
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Minimizing errors in RT-PCR detection and quantification of SARS-CoV-2 RNA for wastewater surveillance
Warish Ahmed1, Stuart L Simpson2, Paul M Bertsch1
1CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, QLD 4102, Australia.
The Science of the Total Environment
|November 25, 2021
Summary
Wastewater surveillance effectively monitors Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) using RT-PCR. This review details strategies to improve accuracy and reliability in detecting SARS-CoV-2 in wastewater, crucial for public health.
Area of Science:
- Environmental microbiology and virology
- Public health surveillance
- Molecular diagnostics
Background:
- Wastewater surveillance using RT-PCR is a cost-effective method for community-level public health monitoring, including COVID-19.
- SARS-CoV-2 wastewater surveillance offers an early warning system for community infections, with rapidly growing laboratory capacity.
- Lack of standardized protocols and QA/QC procedures hinders reliable SARS-CoV-2 wastewater surveillance.
Purpose of the Study:
- To conduct a technical review of factors causing false-positive and false-negative errors in SARS-CoV-2 wastewater surveillance.
- To recommend strategies for identifying and mitigating errors in SARS-CoV-2 RNA detection in wastewater.
- To enhance the accuracy and interpretability of wastewater surveillance data for public health applications.
Main Methods:
- Technical review of existing literature and practices in SARS-CoV-2 wastewater surveillance.
- Analysis of potential sources of error in sample collection, processing, and molecular analysis (RT-PCR).
- Development of recommendations for quality assurance, quality control, and data interpretation.
Main Results:
- Identified key factors contributing to errors in SARS-CoV-2 wastewater surveillance, including sampling, concentration, extraction, and PCR inhibition.
- Proposed stringent QA/QC measures, representative sampling, effective concentration/extraction, PCR inhibition assessment, and process controls.
- Emphasized the need for clear data interpretation guidelines, corrective actions for inconclusive results, and interlaboratory comparisons.
Conclusions:
- Implementing recommended strategies can significantly improve the reliability and accuracy of SARS-CoV-2 wastewater surveillance.
- Wastewater surveillance proved effective during the COVID-19 pandemic and holds future potential for monitoring other communicable diseases.
- Standardization and robust QA/QC are essential for maximizing the utility of wastewater-based epidemiology.
